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 Bioacoustics
The International Journal of Animal Sound and its Recording

ISSN: 0952-4622 (Print) 2165-0586 (Online) Journal homepage: http://www.tandfonline.com/loi/tbio20

A test of the Acoustic Adaptation Hypothesis in

three types of tropical forest: degradation of male
and female Rufous-and-white Wren songs

Brendan A. Graham, Luis Sandoval, Torben Dabelsteen & Daniel J. Mennill

To cite this article: Brendan A. Graham, Luis Sandoval, Torben Dabelsteen & Daniel J.
Mennill (2016): A test of the Acoustic Adaptation Hypothesis in three types of tropical
forest: degradation of male and female Rufous-and-white Wren songs, Bioacoustics, DOI:
10.1080/09524622.2016.1181574

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 Bioacoustics, 2016
http://dx.doi.org/10.1080/09524622.2016.1181574

A test of the Acoustic Adaptation Hypothesis in three types of

tropical forest: degradation of male and female Rufous-and-
white Wren songs
Brendan A. Grahama, Luis Sandovalb, Torben Dabelsteenc and Daniel J. Mennilla
a
Department of Biological Sciences, University of Windsor, Windsor, Canada; bEscuela de Biologa, Universidad
de Costa Rica, San Jos, Costa Rica; cBehavioural Ecology Group, Department of Biology, University of
Copenhagen, Copenhagen, Denmark
Downloaded by [Brendan Graham] at 08:38 13 May 2016

ABSTRACT ARTICLE HISTORY

Many animals produce complex vocalizations that show pronounced Received 18 January 2016
variation between populations. The Acoustic Adaptation Hypothesis Accepted 10 April 2016
helps to explain this variation, suggesting that acoustic signals are
KEYWORDS
optimized for transmission through different environments. Little Acoustic Adaptation
is known about the transmission properties of female vocalizations Hypothesis; bird songs;
because most studies of the Acoustic Adaptation Hypothesis have female song; habitat;
focused on male vocalizations of organisms living at temperate song elements; sound
latitudes. We explored the relationship between environmental transmission
variation and the transmission properties of songs of Rufous-and-
white Wrens, resident Neotropical songbirds where both sexes sing.
Using playback, we broadcast and re-recorded elements of male and
female songs from three populations of wrens living in three different
forest habitats in Costa Rica. We measured four variables of the
re-recorded sounds: signal-to-noise ratio, excess attenuation, tail-to-
signal ratio and blur ratio. Our results show a significant difference
between transmission characteristics of both male and female song
elements across the three habitats, indicating that sounds transmit
differently through different types of tropical forest. The population
from which the broadcast sounds were recorded (source population)
had little effect on sound transmission, however, suggesting that
acoustic differences between these populations may not arise through
acoustic adaptation to these habitats. Male and female elements
showed similar transmission properties overall, although signal-to-
noise ratio of male elements was influenced by source population,
whereas blur ratio and excess attenuation of female elements were
influenced by source population. Our study highlights the differences
in transmission characteristics of animal sounds through different
habitats, and reveals some sex differences in transmission properties.

Introduction
Diverse animal taxa produce long-range acoustic signals that play an important role in mate
attraction and resource defence (Bradbury and Vehrencamp 2011). Animal acoustic signals

CONTACT Brendan A. Graham graham1g@uwindsor.ca

Supplementary material for this article is available via the supplementary tab on the articles online page at
http://dx.doi.10.1080/09524622.2016.1181574
2016 Informa UK Limited, trading as Taylor & Francis Group
 2 B. A. Graham et al.

exhibit incredible diversity, and many signals vary geographically (Marler and Tamura
1964; Irwin et al. 2001; Campbell et al. 2010; Trefry and Hik 2010). Geographic variation
in acoustic signals can play an important role during speciation when different popula-
tions develop divergent acoustic signals and then fail to recognize each other following
secondary contact (Irwin et al. 2001). Given the role that acoustic divergence can play in
evolution, understanding the forces that drive acoustic divergence remains an important
area of research (Wilkins et al. 2013).
Habitat affects the evolution of acoustic signals (Morton 1975; Wiley and Richards 1978;
Hunter and Krebs 1979; Hanford and Lougheed 1991; Dabelsteen et al. 1993; Boncoraglio
and Saino 2007). This widely supported fact lead Morton (1975) to propose the Acoustic
Adaptation Hypothesis: acoustic signals are optimized for transmission through the nat-
ural environment of the animals that produce them, and acoustic signals used for long-
range communication should exhibit adaptations that minimize degradation and maximize
transmission (Morton 1975; Marten et al. 1977; Boncoraglio and Saino 2007). A review by
Boncoraglio and Saino (2007) found that song characteristics of forest and non-forest birds
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vary between habitats, providing further support that the Acoustic Adaptation Hypothesis
may explain acoustic divergence between and within species (Hunter and Krebs 1979;
Tubaro and Segura 1994; Slabbekoorn and Smith 2002). Other studies have found less
support for the Acoustic Adaptation Hypothesis (Rothstein and Fleischer 1987; Date and
Lemon 1993; Daniel and Blumstein 1998; Doutrelant et al. 1999; Trefry and Hik 2010),
although it is noteworthy that a failure to find a relationship between habitat and acoustic
characteristics does not mean that habitat does not affect animals acoustic signals (Barker
2008). In addition to habitat, many other factors influence the evolution of acoustic signals,
including morphology, phylogeny, physiology, sexual selection, social eavesdropping, pred-
ators, learning, founder effects, drift, and other aspects of the environment (e. g. humidity
and ambient noise; Forrest 1994; Lynch 1996). These factors may act in concert and therefore
the evolution of acoustic signals is necessarily complex, and likely to reflect interactions
among these various factors (Forrest 1994; Wilkins et al. 2013).
Dense vegetation can cause significant problems for the transmission of acoustic signals
(Bradbury and Vehrencamp 2011). In particular, leaves, branches, and tree trunks can
degrade signals, changing sounds as they propagate through the environment (Richards
and Wiley 1980; Dabelsteen et al. 1993; Badyaev and Leaf 1997). Degradation is expected
to affect amplitude, frequency composition, and temporal patterns of sounds through pro-
cesses that include scattering, atmospheric turbulence, boundary affects, reverberation, and
dispersion (Richards and Wiley 1980; Dabelsteen et al. 1993; Bradbury and Vehrencamp
2011). Given the important role that habitat plays on the evolution of songs, testing the
transmission properties of an animals acoustic signal through its environment will provide
further insight into the constraints that affect the evolution of signals.
Tropical species present exciting systems for studying the effects of habitat on acoustic
signals, given the high diversity of habitat types, and the dramatic differences in habitats
over relatively short distances (Stutchbury and Morton 2001). Population-level studies of
broadly distributed species are especially revealing, because they provide the opportunity
to examine characteristics of acoustic signals in animals that inhabit a diverse range of
habitats (e.g. Hanford and Lougheed 1991; Slabbekoorn and Smith 2002). The high rates
of philopatry and heightened habitat specialization that are common to many tropical bird
species (Stutchbury and Morton 2008) suggest that tropical animals may be locally adapted
 Bioacoustics 3

to their habitats. Yet, most studies of the Acoustic Adaptation Hypothesis have been con-
ducted on temperate species, and specifically on male song (Barker 2008). Tropical bird
species are interesting from an acoustic perspective, given that females of many tropical
bird species sing (Slater and Mann 2004), an uncommon phenomenon in north-temperate
animals (Price et al. 2009). Studying the acoustic signals of female birds is important (Barker
2008), given that female song is an ancestral trait in birds (Odom et al. 2014), and many
aspects of female song production and development remain poorly understood (Riebel
2003). Comparisons of male and female song characteristics offer a compelling area of
research given that very few geographic-level comparisons have been made between male
and female song characteristics (but see Mennill and Rogers 2006).
To investigate acoustic adaptation across both sexes and among different types of trop-
ical habitats, we studied the transmission properties of songs of Rufous-and-white Wrens
(Thryophilus rufalbus), a year-round resident of Central America and north-western South
America. This species lives in a variety of forested habitats across its range (Stiles and Skutch
1989; Stotz et al. 1996). Interestingly, both male and female Rufous-and-white Wrens sing
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solo songs and produce coordinated duets by combining their solo songs (Mennill and
Vehrencamp 2005). Both males and females possess song repertoires, singing up to 15
different song types (Harris et al. 2016), although male repertoires are larger than female
repertoires (Mennill and Vehrencamp 2005). Male and female songs include similar char-
acteristics, beginning with varied introductory elements, followed by a trill (the longest
part of the song), and usually concluding with a single loud note that is often the highest
frequency part of the song (Mennill and Vehrencamp 2005). Given that both sexes sing
within this species, this system allows us to compare patterns between sexes and further
our understanding of female song.
We used recordings of played-back songs to examine the transmission properties of both
male and female Rufous-and-white Wren songs in three different populations in Costa Rica.
Previous work has demonstrated that songs of Rufous-and-white Wrens vary geographically
(Valderrama et al. 2007), and our ongoing research confirm that songs are variable between
our three study populations (based on fine structural measurements, i.e. syllable length,
bandwidth and dominant frequency of the trills). Our three study sites vary in habitat
structure, vegetation density, and climate (Clark et al. 2002; Mata and Echeverria 2004), and
therefore acoustic differences may reflect local adaptations at each site. We sought to test
whether variation between songs among populations shows evidence of acoustic adaptation.
Specifically, we explored the relationship between habitat and acoustic structure of male
and female Rufous-and-white Wren songs, testing whether sound propagation varied with
playback site (i.e. the location where sounds were broadcast and re-recorded) and source
population (i.e. the location where the stimuli were recorded).

Methods
Study site
We conducted our experiment at three sites in Costa Rica: Sector Santa Rosa of the
Guanacaste Conservation Area (10.8836N, 85.7750W, 300m a.s.l.); Sector Rincon de la
Vieja of the Guanacaste Conservation Area (10.8300N, 85.3239W, 1000m a.s.l.); and the
San Luis Valley of Monteverde at the University of Georgia Costa Rica field site (10.2380N,
 4 B. A. Graham et al.

84.7970W, 1100m a.s.l.). Populations of free-living Rufous-and-white Wrens are found at

all three sites. Playback sessions took place in 2013 on April 1718 at San Luis, June 12 at
Rincon de la Vieja, and June 1112 at Santa Rosa during the onset of the breeding season
at each population (birds breed earlier at San Luis than the other two sites; pers. obs.). All
playback sessions were conducted between 0700 and 1100h, a time period when this species
is most vocally active (Mennill and Vehrencamp 2005). We conducted our experiment over
a two-day period at each site, to ensure that weather conditions like temperature, relative
humidity and wind were consistent throughout the experiment. Daily temperatures were
consistent with mean monthly values at each of the sites (average temperature and relative
humidity ranged from 23.0C and 72.6% at the montane forest site, 26.0C and 84.0% at the
wet site and 27.3C and 76.0% at the dry forest site over the two day periods), and therefore
we feel confident that the meteorological conditions are representative of conditions at each
site during the appropriate time of year.
Our three study sites differ in both vegetation and precipitation (Clark et al. 2002; Mata
and Echeverria 2004). (1) Santa Rosa (hereafter referred to as the dry forest site) is a trop-
Downloaded by [Brendan Graham] at 08:38 13 May 2016

ical dry forest (following the Holdridge Life Zone classification system, Holdridge 1967)
with a dry season that lasts from November to April and an intense rainy season from
May to November (1876mm on average/year from 1998 to 2013; NASA TRMM project).
The understory at this dry forest site is relatively open (basal area = 25.0 m2 Ha1 for
stems>10cm; Gillespie et al. 2000) especially during the dry season, when the majority of
shrubs in the understory are leafless. Vegetation density increases following the start of the
rainy season. The canopy attains heights of approximately 20m although some emergent
trees reach heights of 30m (Janzen 1983). (2) Rincon de la Vieja (hereafter referred to as
the wet forest site) is a Premontane Moist-Wet Forest (Holdridge 1967), with a dry season
from January to April (2057mm average/year from 1998 to 2013; NASA TRMM project).
This area is wetter than the lowland dry forest, but receives less precipitation than forests at
higher elevations. This forest type is representative of many mid-elevation forests (~900m
elevation); the understory is relatively open, with fewer shrubs found here than in the dry
forest (basal area=31.2m2Ha1; Heaney and Proctor 1990; basal area data are not available
from our wet forest site, and this value is chosen for a comparison site in Costa Rica with
similar vegetation, climate and altitude). The canopy attains heights of 2530m, and many
large trees, including figs, dominate the forest (Janzen 1983). (3) San Luis field station at
Monteverde (hereafter referred to as the montane forest site) is a Lower Montane Wet
Forest (~1100m elevation; Holdridge 1967), with a season of less precipitation lasting from
January to April. This area receives greater precipitation than our other two sites (2706mm
average/year from 1998 to 2013; NASA TRMM project). The understory is densely vege-
tated by shrubs, ferns and palms (basal area=62.0m2 Ha1 for stems>10cm; Nadkarni
et al. 1995) with epiphytes covering 5070% of the tree trunks. Consequently, this habitat is
much more dense than the understory at our other two sites (Janzen 1983). The canopy at
the montane forest site reaches heights of 2530m, dominated by diverse large tree species.

Song type selection

For our playback stimuli, we used both male and female songs that we recorded from each
of the three study populations in 2012. Recordings were collected using a solid-state digi-
tal recorder (PMD-660 Marantz; 44.1kHz sampling rate; 16-bit accuracy; WAVE format)
 Bioacoustics 5

and a shotgun microphone (Sennheiser MKH70). To create our stimuli, we chose five
of our highest quality songs from each population for each sex (each song used for the
stimuli came from a different individual), using only songs with high signal-to-noise ratio
(assessed visually based on sound spectrograms) and no overlap from other conspecific
or heterospecific sounds. From those songs, we selected population-specific elements that
were representative of elements that were most common in each population during our
recording sessions. To create our final playback stimuli, we selected 18 male song elements
(6 from each population, giving rise to 6 introductory, trill and terminal syllables overall;
Figure 1) and 20 female song elements (6 from the montane and dry forest sites and 8 from
the wet forest site, giving rise to 7 introductory, and terminal syllables, and 6 trill syllables,
overall; we included 2 additional elements for wet forest females to reflect the diversity of
female song elements in that population; Figure 1). We determined that six elements from
each sex at each population was an appropriate number, given that the elements we selected
for both male and female playback are representative of elements that are widespread and
frequently used within each population. Our sample size (n=18 elements for males and
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n=20 elements for females) is comparable to previous transmission studies of species with
intermediate to large song repertoires (Holland et al. 1998; Barker et al. 2009; Mockford
et al. 2011). We isolated and filtered songs and elements using the FFT filter function of
Audition software (version 3.0, Adobe Systems, San Jose, CA, USA); for each sound, we

4
male songs female songs
3

1
D W M D W M
0
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18
4
male introductory elements male trill elements male terminal elements
Frequency (kHz)

1
D D W W M M D D W W M M D D W W M M
0
0 1 2 3 4 5 6 7
4
female introductory elements female trill elements female terminal elements
3

1
D D W W W M M D D W W M M D D W W W M M
0
0 1 2 3 4 5 6 7
Time (sec)

Figure 1. Sound spectrograms of example male and female Rufous-and-White Wren songs recorded
from each of the three populations where playback experiments were conducted (top row). Sound
spectrograms of example male song elements (second row) and female song elements (third row) used
for playback during the transmission experiment. Letters indicate the population where the song or song
element was recorded (D=dry forest, W=wet forest and M=montane forest).
 6 B. A. Graham et al.

used a different filter (see page 1 of supplementary material for information on the filters
used to isolate each sound), given that each sound occupied a different bandwidth.
We focused our analysis on elements within the male and female songs, rather than entire
songs, because we were interested in understanding how the degradation of single elements
contributes to the degradation of entire songs. Examining elements separately from entire
songs is important, given that the context in which sounds are broadcast can affect the
acoustic properties; for example, reverberation is known to enhance both the length and
amplitude of a sound, especially for the pure tone elements used by many forest birds, that
change little in frequency (Slabbekoorn et al. 2002; Nemeth et al. 2006). While we present
the results for elements only in this manuscript, we did analyse entire songs in another
analysis, and we found that songs showed a similar pattern to elements (see supplementary
material Tables S3S7).
Using these prepared sounds, we created playback tracks by pasting the stimuli into a
single file using Audition (Adobe Systems Inc., San Jose, CA). Each stimulus track included
5.0s of silence at the outset (facilitating a measurement of background noise), followed
Downloaded by [Brendan Graham] at 08:38 13 May 2016

by each of the sounds in succession, with 1.5s of silence between each sound (preventing
sounds from being overlapped by the end of the previous sound). Each playback stimulus
was played five times in succession to maximize the chances of recording multiple examples
of each element without overlap from background sounds. Each repetition was separated
by 5.0s of silence before the next repetition began.

Experimental set-up
At each of our three sites, we conducted our transmission experiment in three different
Rufous-and-white Wren territories. We chose territories that were representative of the
common vegetation at each site. Within each territory, we positioned both the speaker and
microphone at a single height above ground (1.5m). This height falls within the range of
perch heights (1 to 5m) male and female Rufous-and-white Wrens are most commonly
observed using as song posts (Barker and Mennill 2009). We placed the microphone at
four separate distances (5, 10, 20 and 40 m) from the speaker. We chose 20 m as one
important distance based on a previous microphone array study that found 20m to be the
average distance separating male and female Rufous-and-white Wrens while performing
duets (Mennill and Vehrencamp 2008). The maximum (40m) and minimum (5 and 10m)
distances were chosen based on doubling and halving this average distance. Unlike previous
studies (e.g. Barker et al. 2009; Sabatini et al. 2011), where playback was conducted along a
linear transect, we distributed the four distances at different axes within each territory (as
in Sandoval et al. 2015). By doing this, we attempted to include more of the birds territo-
ries in our transmission tests, thus providing a more representative sampling of the effect
of habitat on sound transmission. We chose these playback axes according to the cardinal
points in all of the nine territories where we conducted our playback.
We broadcast sounds using an active loudspeaker (Anchor Audio, Minivox; frequency
response 0.112 kHz), and re-recorded them using an omnidirectional microphone
(Sennheiser ME62) and a solid-state recorder (PMD-660 Marantz; 44.1 kHz sampling
rate; 16-bit accuracy; WAVE format), connected to a pre-amplifier (Sound Device MP-1:
frequency response 0.0222kHz). Playback was broadcast at 75dB (as measured at 1m dis-
tance using a sound meter; Radio Shack model 332055 using C-weighting slow response),
 Bioacoustics 7

allowing us to match the sound pressure level that has been used in a previous study of
Rufous-and-white Wrens songs (Barker et al. 2009). We increased the gain on our pre-
amplifier to 18 and 28dB for the 20- and 40-m trials, respectively, and we correct for these
changes in gain by adding 18 and 28 dB to the appropriate analyses. Changing the gain was
a critical component of these recordings, because the same recording levels could not be
used to collect high-quality recordings for both the short and long transmission distances.

Sound analyses
As in most other transmission studies (e.g. Holland et al. 1998; Lampe et al. 2007; Barker
et al. 2009), we used SigPro software (v 3.25; Pedersen 1998) to analyse the transmission
properties of all recorded sounds. We compared recorded sounds at the four distances (5, 10,
20 and 40m) against a model signal. The model signal used for comparison was obtained by
broadcasting our male and female stimuli with the aforementioned playback and recording
apparatuses, but with a separation distance of just 1.25m at a height of 1.5m on a flat dirt
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road in Sector Santa Rosa i.e. an environment with no vegetation (in a 20-m radius) that
could influence the transmission between the speaker and the microphone on a calm
morning with little or no background noise (e.g. wind). We then filtered and trimmed these
recordings for the purpose of removing any potential tails or echoes introduced during
the model signal recording. We used these model signals, rather than the original stimuli,
to account for any noise that might have been introduced by the playback or recording
equipment (as in Lampe et al. 2007, for example).
We compared degraded sounds to model sounds to obtain four measurements of deg-
radation (for details see Dabelsteen et al. 1993; Holland et al. 2001): signal-to-noise ratio,
tail-to-signal ratio, blur ratio and excess attenuation. We also measured background noise by
sampling the background sound immediately prior to each stimulus recording (as described
by Dabelsteen et al. 1993). We assumed that this background sound matched the noise over-
lapping our re-recorded playback sounds (Holland et al. 1998; Barker et al. 2009; Sabatini
et al. 2011). Background noise was filtered within the same frequency ranges as the test
sounds and then used to calculate signal-to-noise ratio and better understand how signal-
to-noise ratio varied among our three forested sites, as described in Dabelsteen et al. 1993.
Furthermore, we measured and compared background noise at each area, so that we could
quantify the level of environmental noise at each site for each sound within its frequency
range, given that past studies have shown that the background noise varies with frequency,
that there are differences in the amount of ambient noise between forested habitats, and
that these differences can affect sound degradation (Slabbekoorn et al. 2002).
For each sound, we analysed up to three re-recorded exemplars per distance along each
transect, although in some instances, we were unable to measure three exemplars due to
overlap by background noise. Due to windy conditions at Monteverde, we were only able
to collect useful measurements for two of the three transects at 5, 10 and 20m and only
one of the three transects at 40m; the remaining sounds were too heavily overlapped by
background noise. After omitting these overlapped sounds, we were left with 1600 meas-
urements for male song elements (2.471.00 per distance in each transect; meanSE),
and 1770 for female song elements (2.461.01).
 8 B. A. Graham et al.

Statistical analyses
To analyse degradation of Rufous-and-white Wren sounds, we used linear mixed models.
We analysed the sexes independently with separate models. We used the four sound deg-
radation measurements (signal-to-noise ratio, blur ratio, tail-to-signal ratio and excess
attenuation) as our response variables and ran each of the measurements in a model for each
of the sexes (i.e. eight models in total). For each model, we had four independent variables:
playback site (three levels corresponding to the three sites where we conducted playback),
source population (three levels corresponding to the three populations where birds were
recorded), distance (four levels, corresponding to the four distances between loudspeaker
and microphone) and element type (three levels, because we were interested in seeing if there
were differences in the degradation of introductory, trill and terminal elements; Mennill and
Vehrencamp 2005). For our analysis, we examined main effects and two-way interactions
for each model. We used Tukey post hoc tests to evaluate whether differences in means were
significant. To analyse background noise (dB) during the transmission experiments, we ran
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two additional models, one for each sex. Like our models for sound degradation, we had
four independent variables (playback site, source population, element type and distance),
but for our background noise analysis, we examined only main effects.
To understand whether Rufous-and-white Wrens song elements show local adaptation
to the environment where the birds are found, we focused on the interaction playback
sitesource population. We focused specifically on this interaction, based on our expec-
tation that elements that are adapted to their local environment should transmit more
effectively (i.e. experience less degradation) at the playback site where they were originally
recorded.
We report all values as meanSE. All analyses were performed in JMP (version 10.0;
SAS Institute, Cary, NC, USA).

Results
Our transmission data reveal that playback site and source population had different effects
on the degradation of male and female Rufous-and-white Wren song elements; transmis-
sion properties regularly showed a significant effect of playback site, but rarely showed a
significant effect of source population. Below, we present detailed findings for male and
then female song elements, describing the main effects followed by the interaction terms.

Males
For male song elements, signal-to-noise ratio, tail-to-signal ratio and excess attenuation
were all significantly affected by playback site (Table 1); signal-to-noise ratio was higher at
the wet and dry forest sites than at the montane forest site, tail-to-signal ratio was higher
at the dry forest site than the other two sites, and excess attenuation was greater at both
the wet and dry forest sites than the montane forest site (Figure 2). Signal-to-noise ratio
was the only measurement that was significantly affected by source population (Table 1);
elements recorded from the montane and wet forest sites had a higher signal-to-noise
ratio than elements recorded from the dry forest site (Figure 3). All four sound degrada-
tion measurements were significantly affected by distance (Table 1); degradation increased
 Downloaded by [Brendan Graham] at 08:38 13 May 2016

Table 1.Main effects and two-factor interactions for linear mixed models analysing male song elements for each of four measures of degradation of Rufous-and-white
Wren song elements.
Signal-to-noise ratio Tail-to-signal ratio Blur-ratio Excess attenuation
Male elements df F p df F p df F p df F p
Model 39 109.18 <0.001 39 23.61 <0.001 39 7.1 <0.001 39 75.02 <0.001
Playback site 2 61.03 <0.001 2 15.78 <0.001 2 0.12 0.889 2 81.17 <0.001
Source population 2 11.33 <0.001 2 1.33 0.266 2 1.99 0.137 2 0.03 0.974
Distance 3 967.13 <0.001 3 222.03 <0.001 3 24.05 <0.001 3 749.43 <0.001
Element type 2 8.70 <0.001 2 2.77 0.063 2 3.58 0.028 2 13.48 <0.001
Playback sitesource population 4 3.67 <0.001 4 0.17 0.951 4 0.53 0.715 4 2.30 0.057
Playback sitedistance 6 36.88 <0.001 6 4.72 <0.001 6 2.36 0.029 6 37.36 <0.001
Playback siteelement type 4 12.74 <0.001 4 1.65 0.158 4 2.29 0.058 4 2.37 0.051
Source populationdistance 6 1.53 0.165 6 0.99 0.432 6 0.68 0.665 6 1.26 0.275
Source populationelement type 4 21.35 <0.001 4 11.45 <0.001 4 4.85 0.001 4 4.67 0.001
Distanceelement type 6 2.27 0.034 6 0.46 0.838 6 3.22 0.004 6 2.01 0.062
The significance of bold values is (p < 0.05).
Bioacoustics
9
 10 B. A. Graham et al.

Males Females
50 50
Signal-to-noise
ratio (dB) 40 40

30 30

20 20

10 10
a b b a b c
0 0
-25 -25
Tail-to-signal ratio

-20 -20

-15 -15
(dB)

-10 -10

-5 -5
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a a b a a b
0 0
0.08 0.06

0.06
0.04
ratio
Blur

0.04
0.02
0.02

a a a a a b
0 0
30 25
attenuation (dB)

20
20
Excess

15

10
10
5
a b b a b b
0 0
Montane Wet Dry Montane Wet Dry
forest forest forest forest forest forest
Playback site

Figure 2.Four measurements of sound degradation of rufous-and-white wren song elements at each of
three different playback sites in Costa Rica, both for males (left column) and females (right column). Error
bars are standard errors of the mean, and bars with different letters indicate that values are significantly
different from each other in post hoc tests.

as distance from the speaker increased (Table S1). Three of the four sound degradation
measurements (signal-to-noise ratio, blur ratio and excess attenuation) showed significant
variation with element type (Table 1); signal-to-noise ratio was higher for introductory
and terminal elements than trill elements, blur ratio was higher for terminal elements than
either introductory or trill elements, and excess attenuation was higher for introductory
elements, than either terminal or trill elements (Figure 4).
 Bioacoustics 11

Males Females
50 40
Signal-to-noise
40
ratio (dB) 30
30
20
20
10
10
a a b a a a
0 0
-20 -20
Tail-to-signal ratio

-15 -15
(dB)

-10 -10

-5 -5
Downloaded by [Brendan Graham] at 08:38 13 May 2016

a a a a a a
0 0
0.10 0.06

0.08
0.04
0.06
ratio
Blur

0.04
0.02
0.02
a a a a b b
0 0
25 30
attenuation (dB)

20
20
Excess

15

10
10
5
a a a a b a
0 0
Montane Wet Dry Montane Wet Dry
forest forest forest forest forest forest

Source Population

Figure 3.Four measurements of sound degradation of rufous-and-white wren song elements based on
the source population (where a sound was recorded), both for males (left column) and females (right
column). Error bars are standard errors of the mean, and bars with different letters indicate that values
are significantly different from each other in post hoc tests.

All four sound degradation measurements showed significant interaction effects in our
analysis of male song elements, especially for the interactions between playback sitedis-
tance (Table S1) and source populationelement type (Table 1). Signal-to-noise ratio of
elements for the interaction playback sitedistance was significantly higher at shorter
distances (both 5 and 10m) at the wet and dry forest sites, and lowest at the furthest dis-
tances (20 and 40m) at the montane forest site (Table S1). Like the patterns observed for
signal-to-noise ratio, tail-to-signal and blur ratio were higher for elements at the furthest
 12 B. A. Graham et al.

Males Females
50 50
Signal-to-noise 40 40
ratio (dB)
30 30

20 20

10 10
a a b a b c
0 0
-20 -25
Tail-to-signal ratio

-20
-15
-15
(dB)

-10
-10
-5
-5
Downloaded by [Brendan Graham] at 08:38 13 May 2016

a a a a b c
0 0
0.08 0.08

0.06 0.06
ratio
Blur

0.04 0.04

0.02 0.02

a,b a b a b c
0 0
25 30
attenuation (dB)

20
20
Excess

15

10
10
5
a b b a a a
0 0
Introductory Trill Terminal Introductory Trill Terminal

Element Type

Figure 4.Four measurements of sound degradation of rufous-and-white wren song elements based on
element type for males (left column) and females (right column). Error bars are standard errors of the
mean, and bars with different letters indicate that values are significantly different from each other in
post hoc tests.

distances at all three sites, while excess attenuation was greatest at the furthest distances at
the wet and dry forest sites, with the lowest values being observed at the shortest distances
at the montane forest site. For the interaction between source populationelement type,
the majority of element types recorded from both montane and wet forest sites had a higher
signal-to-noise ratio than element types recorded from our dry forest site (Table S2). Tail-
to-signal ratio was lower for terminal and introductory elements from the montane and
wet forest sites, while tail-to-signal ratio was highest for trill elements recorded from the
 Bioacoustics 13

50

Signal-to-noise 40
ratio (dB)
30

20

10
a,b a a c c d,e c,d c,d b,e
0
50
Signal-to-noise

40
ratio (dB)

30

20

10
a,b a b c c c d d d
Downloaded by [Brendan Graham] at 08:38 13 May 2016

0
Montane Wet Dry Montane Wet Dry Montane Wet Dry
forest forest forest forest forest forest forest forest forest

Montane forest Wet forest Dry forest

Figure 5.Signal-to-noise ratio measurements of Rufous-and-white Wren song elements showing the
interaction of playback sitesource population for males (top) and females (bottom). Error bars are
standard errors of the mean, and bars with different letters indicate that values are significantly different
from each other in post hoc tests.

montane and wet forest sites and introductory and terminal elements recorded from the dry
forest site. Wet forest terminal and trill elements along with dry forest terminal elements
showed a lower blur ratio than dry forest trill elements. Finally, excess attenuation was sig-
nificantly higher for montane and wet forest introductory elements than trill or terminal
elements from the same populations (Table S2). Only a single interaction (signal-to-noise
ratio) was significant for the interaction playback sitesource population; however, sounds
did not show significantly less degradation at the sites where they were recorded (i.e. the
degradation of elements recorded at the dry forest was not lower than elements recorded
at our wet and montane forest sites, when played at our dry forest site; Figure 5). Elements
recorded from montane and wet forest sites had a higher signal-to-noise ratio at the wet and
dry forest sites, while signal-to-noise ratio of elements (from all three populations) played
at the montane forest site had the lowest signal-to-noise ratio values. Signal-to-noise ratio
was the only variable to show a significant relationship for the interaction between playback
siteelement type, where introductory and trill elements played at the montane forest site
had the lowest signal-to-noise ratio values from all others (Table S1). There were no sig-
nificant effects for the interaction source populationdistance, while distanceelement
type affected signal-to-noise ratio and blur ratio only. For signal-to-noise ratio, elements at
the closest distances (5m) had a higher signal-to-noise ratio than elements at the farthest
distances (40m). Meanwhile terminal elements at farther distances (20 and 40m) had a
significantly higher blur ratio than all other element types, while trill and introductory
elements at shorter distances (5 and 10m) had the lowest blur ratio values (Table S1).
 Downloaded by [Brendan Graham] at 08:38 13 May 2016

14
B. A. Graham et al.

Table 2.Main effects and two-factor interactions for linear mixed models analysing female song elements for each of four measures of degradation of Rufous-and-
white Wren song elements.
Signal-to-noise ratio Tail-to-signal ratio Blur-ratio Excess attenuation
Female elements df F p df F p df F p df F p
Model 39 126.55 <0.001 39 80.45 <0.001 39 32.29 <0.001 39 58.93 <0.001
Playback site 2 84.78 <0.001 2 27.36 <0.001 2 8.48 <0.001 2 50.30 <0.001
Source population 2 0.27 0.763 2 3.09 0.046 2 9.59 <0.001 2 7.83 <0.001
Distance 3 1073.53 <0.001 3 527.32 <0.001 3 88.86 <0.001 3 574.27 <0.001
Element type 2 25.99 <0.001 2 59.99 <0.001 2 24.71 <0.001 2 0.99 0.373
Playback sitesource population 4 5.78 <0.001 4 1.69 0.151 4 1.74 0.140 4 0.60 0.660
Playback sitedistance 6 28.53 <0.001 6 9.20 <0.001 6 8.17 <0.001 6 12.67 <0.001
Playback siteelement type 4 15.12 <0.001 4 0.93 0.448 4 1.57 0.179 4 1.10 0.356
Source populationdistance 6 0.51 0.804 6 1.09 0.368 6 5.57 <0.001 6 1.26 0.274
Source populationelement type 4 33.29 <0.001 4 242.51 <0.001 4 68.90 <0.001 4 1.06 0.374
Distanceelement type 6 1.54 0.162 6 2.33 0.030 6 4.21 <0.001 6 1.11 0.355
The significance of bold values is (p < 0.05).
 Bioacoustics 15

Females
For female song elements, sound degradation was significantly affected by the majority of
the main effects (Table 2). Signal-to-noise ratio, tail-to-signal ratio, blur ratio and excess
attenuation were all significantly affected by playback site. Female elements showed a higher
signal-to-noise ratio and tail-to-signal ratio, lower blur ratio, and experienced greater excess
attenuation at the dry forest site, while elements played at the montane forest site exhibited
a lower signal-to-noise ratio and tail-to-signal ratio, higher blur ratio, but experienced less
excess attenuation (Figure 2). Source population affected tail-to-signal ratio, blur ratio and
excess attenuation (Table 1). While post hoc tests revealed no differences among sites for
tail-to-signal ratio, female elements recorded from the montane forest site had a lower blur
ratio than elements from the other two populations; elements recorded from the dry and
montane forest showed greater excess attenuation than elements recorded from the wet
forest (Figure 3). Like male elements, all four measurements were affected by distance, and
elements showed greater degradation at the furthest distances (Table S2). Lastly, three of the
Downloaded by [Brendan Graham] at 08:38 13 May 2016

four measurements (signal-to-noise ratio, tail-to-signal ratio and blur ratio) were affected
by element type (Table 2), and terminal elements had a higher signal-to-noise ratio, higher
tail-to-signal ratio and higher blur ratio than both introductory and trill elements (Figure 4).
Half of the interactions showed significant effects in our analysis of female song ele-
ments (Table 2). Signal-to-noise ratio was the only measurement that showed a significant
pattern for playback sitesource population, where elements had a significantly higher
signal-to-noise ratio when played at our dry forest site than at our montane and wet forest
sites, and elements played at wet forest site had a significantly higher signal-to-noise ratio
than elements at our montane forest site (Figure 5). However, as we observed for males,
degradation of non-local elements was not significantly greater than that of local elements
outside of the populations where they were recorded. All four degradation measurements
were significant for the interaction playback sitedistance (Table S2); song elements expe-
rienced significantly greater degradation as distance from the speaker increased, similar to
patterns observed for males. Signal-to-noise ratio was significant for playback siteelement
type (Table S2), and elements had a significantly higher signal-to-noise ratio at the dry
forest site, followed by the wet and montane forest sites (Table S2). Only blur ratio showed
a significant effect for the interaction between source populationdistance; elements from
the wet and dry forest sites at the furthest distances had a higher blur ratio than elements
from the montane forest site at all distances (Table S2). Signal-to-noise ratio, tail-to-signal
ratio, and blur ratio were significant for source populationelement type, where signal-to-
noise ratio was significantly lower for trill elements from all populations than the majority
of terminal and introductory elements (Table S2). Tail-to-signal ratio was lower for ter-
minal elements recorded from our montane forest site, while introductory elements from
our montane forest site have the longest tails. Greater blur ratio was exhibited by terminal
elements from the dry and wet forest sites than introductory and trill elements (Table S2).
Finally, blur ratio was the only measurement significant for element type x distance, and
revealed that terminal and introductory elements at the furthest distances (20 and 40m)
experienced a higher blur ratio than trill elements at all distances (Table S2).
 16 B. A. Graham et al.

Table 3.Main effects and two-factor interactions for the linear mixed models analysing comparisons of
background noise during male and female song elements.
Male song elements Female song elements
df F p df F p
Model 39 26.02 <0.001 12 32.92 <0.001
Playback site 2 6.99 0.001 2 7.47 0.001
Source population 2 0.11 0.899 2 0.20 0.822
Distance 3 155.39 <0.001 3 189.64 <0.001
Element type 2 1.71 0.182 2 1.87 0.155

Background noise
Transmission experiments for both male and female song elements showed that background
noise varied by site (Table 3). Background noise at the montane forest site was significantly
higher than at the wet and dry forest sites, which were not significantly different from one
another (Table S7). Source population did not show a significant effect for background
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noise levels for either male or female elements (Table 3), while distance significantly affected
both male and female songs, where background noise increased with distance between the
loudspeaker and the microphone (Table S7).

Discussion
Using a sound-transmission experiment, we tested the influence of habitat on the trans-
mission of male and female Rufous-and-white Wren song elements in three different types
of tropical forest, thereby testing predictions of the Acoustic Adaptation Hypothesis. We
found that playback site affects the transmission of both male and female elements, and we
found significant differences in background noise levels among sites. Source population (i.e.
the location where songs were recorded) had little effect on degradation, given that only
four of eight degradation measurements were significant (i.e. signal-to-noise ratio for male
elements and tail-to-signal ratio, blur ratio and excess attenuation for female elements).
Furthermore, the interaction playback sitesource population did not suggest that song
elements are locally adapted, given that elements did not experience less degradation at
their respective sites (e.g. dry forest song elements did not experience less degradation at
the dry forest site in comparison to elements recorded at our wet or montane forest sites;
Figure 5). Overall, Rufous-and-white wren songs appear to be optimized for transmission
through forested habitat in comparison to open habitats (Barker et al. 2009), but our data
reveal that their song elements are not specifically adapted for transmission through dif-
ferent types of tropical forests. We conclude that habitat influences sound transmission of
both male and female songs, but that sounds in these three study populations do not show
strong evidence of acoustic adaptation to the three different habitats.

Playback site
The Acoustic Adaptation Hypothesis predicts that the signals of animals living in densely
vegetated habitats should be adapted for transmission through these habitats (Richards and
Wiley 1980; Badyaev and Leaf 1997). Support for the Acoustic Adaptation Hypothesis is
mixed (Ey and Fisher 2009); many studies have demonstrated support for the hypothesis
 Bioacoustics 17

(Hunter and Krebs 1979; Tubaro and Segura 1994; Perla and Slobodchikoff 2002; van
Dongen and Mulder 2006; Derryberry 2009), whereas other studies have failed to show
support (Rothstein and Fleischer 1987; Date and Lemon 1993; Daniel and Blumstein 1998;
Doutrelant et al. 1999; Trefry and Hik 2010). In our study, we found that playback site had a
significant effect on the degradation of both male and female acoustic signals. Environmental
differences such as vegetation density, atmospheric absorption and ambient noise all affect
sound transmission (Brumm and Naguib 2009), and differences in these factors between
our three sites surely played a role in the transmission properties we described. We observed
greater degradation at the montane and wet forest sites than at the dry forest site with
regards to tail-to-signal ratio and blur ratio of both male and female elements. Vegetation
density and rainfall are higher at the montane and wet forest sites than the dry forest site,
where the habitat is more open (Nadkarni et al. 1995; Gillespie et al. 2000). Densely for-
ested habitats result in greater degradation because there are more leaves, stems, branches,
and trunks, thereby increasing the effect of reflection, refraction, and diffraction on sound
waves (Naguib 2003).
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In contrast to the pattern for tail-to-signal ratio and blur ratio, excess attenuation was
significantly lower at the montane forest site than at the wet and dry forest sites, for both
male and female elements. While vegetation density does affect excess attenuation, other
factors such as atmospheric scattering and turbulence, as well as boundary interference,
also affect attenuation (Brumm and Naguib 2009). Humidity and temperature are known
to affect the attenuation of sounds, and sounds experience less attenuation in humid air
and when temperatures are cooler (Ingard 1953; Griffin 1971). Among the three study sites,
the montane forest site receives the largest amount of annual rainfall; humidity is greater
(an average of 91% throughout the year; Johnson et al. 2005) and temperatures are cooler
(mean=20.7C; www.worldclim.org) than at the other two sites (by comparison the average
humidity in the dry forest ranges from 20 to 60% during the dry season and temperatures
are warmer; mean=24.8C; Janzen 1988; Clark et al. 2002; Mata and Echeverria 2004).
Therefore, climate differences among sites may contribute to the differences in excess attenu-
ation we observed, as has been suggested in the previous studies (Morton 1975; Nottebohm
1975), although we are aware of no studies that have tested the effect of climate differences
between sites on sound transmission.
Signal-to-noise ratio of both male and female elements was significantly higher when
sounds were played at the wet and dry forest sites than at the montane forest site. These
differences may be attributable to the much noisier environment at our montane forest site,
an idea that was directly supported by our comparisons of background noise (Table S7).
Conditions at the montane forest site were much windier than at the other two sites and
wind produces low-frequency noise in the range of 0.11.0kHz (Bradbury and Vehrencamp
2011). The added background noise masked some of the elements used for playback during
our experiment, especially those produced around 1.0kHz (e.g. the introductory and trill
elements of many male and some female songs are produced at this frequency; Mennill and
Vehrencamp 2005). Additionally, within highland tropical forests, there is a considerable
amount of background noise in the high-frequency spectrum (Ryan and Brenowitz 1985;
Slabbekoorn and Smith 2002). Animals like cicadas call continuously, with this noise band
beginning around 2kHz and extending up to 5kHz (Slabbekoorn 2004). A recent study
by Hart et al. (2015) found that birds avoided temporal overlap with cicadas, suggesting
that biotic noise (from sources including cicadas) may influence the frequency and timing
 18 B. A. Graham et al.

of avian vocal signals. Many of the female elements and songs recorded and used for this
experiment are produced at 3 kHz. Since these sounds fall within the range of high-
frequency noise, female sounds are at risk of being masked by cicada advertising calls, and
background noise differences between sites may explain why we observed a higher signal-
to-noise ratio for female sounds played at the dry forest site (Slabbekoorn 2004).

Source population
Source population had little influence on the degradation of male or female Rufous-
and-white Wren elements. Only male elements showed a significant effect of source popula-
tion for signal-to-noise ratio, where male elements recorded at the montane forest and wet
forest sites showed less degradation than elements recorded at our dry forest site (i.e. higher
signal-to-noise ratios), but for no other degradation measurements. Many animals increase
signal-to-noise ratio to compensate for noisy environments (Brumm and Slabbekoorn
2005). For instance, abiotic features such as wind and fast-flowing rivers produce low-
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frequency noise (0.11.0kHz for wind noise, up to 4kHz for aquatic noise, Slabbekoorn
2004; Bradbury and Vehrencamp 2011) that can mask signals in this range. Background
noise differences among sites likely contributed to the higher signal-to-noise ratio observed
for male elements from the wet and montane forest sites. For example, species living next to
water produce vocalizations at higher frequencies to avoid having their vocalizations masked
by the noise produced by streams (Martens and Geduldig 1990). By comparison, there is
less low-frequency ambient noise at the dry forest site during the breeding season, where
there is little or no moving water, and conditions are less windy. The reduced background
noise may explain why broadband elements are commonly used in songs at the dry forest
site where males often terminate songs using broadband elements (e. g. the second male
terminal element in the second row of Figure 1; 17 of 40 of song types recorded in 201213
included broadband terminal elements). By comparison, male elements (especially terminal
elements, e.g. the fourth and fifth male terminal elements in the second row of Figure 1)
from our wet and montane forest sites tend to be more tonal (Figure 1; only 2 of 35 song
types at our wet forest site, while only 8 of 33 song types at our montane forest site included
broadband terminal elements), suggesting that males use these elements over broadband
signals because they are masked less easily by ambient noise. Differences in signal-to-noise
ratio of elements for male Rufous-and-white Wrens could be indicative of local adaptation,
but could also represent phenotypic plasticity. For instance, Red-wing Blackbirds (Agelaius
phoenicius) make short-term modifications to their songs by increasing their signal tonality
when exposed to low-frequency white noise (Hanna et al. 2011). Evidence from this study
and others (Slabbekoorn and Peet 2003; Mockford et al. 2011; Parris and McCarthy 2013;
Gough et al. 2014) have demonstrated the high plasticity in birds that learn their songs,
where individuals are able to modify their songs in the presence of increased noise to stand
out in their environment.
Female elements did not show differences in signal-to-noise ratio, in contrast to the
pattern observed for males. However, we did observe significant differences for the other
three degradation measurements; these results may indicate local adaptations for female
elements. Tail-to-signal ratio was significant in our overall model, but did not show any
differences among populations. Female elements from the montane forest site had a lower
blur ratio overall than elements recorded from the other two sites. This is likely due to the
 Bioacoustics 19

fact that vegetation density is higher at the montane forest site, suggesting that female ele-
ments from this population are adapted for transmission through dense vegetation. Finally,
we found small differences for the excess attenuation of female elements, with sounds
recorded from the wet forest site showing less excess attenuation than sounds recorded from
the montane and dry forest sites. These differences may be indicative of local adaptation,
given that excess attenuation was highest at the wet forest site (although not significantly
different than excess attenuation at our dry forest site). This result aligns with predictions
of the Acoustic Adaptation Hypothesis, given that we would expect sounds from each of
the three sites to be optimized to their respective sites.

Element type
Both male and female elements showed similar degradation patterns, suggesting that song
elements have evolved under similar influences for both sexes. Nevertheless, it is worthwhile
to note that degradation was not equal across all element types. For example, trill elements
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exhibited a lower signal-to-noise ratio (possibly because they are produced at lower frequen-
cies than other elements and therefore more likely to be masked by background noise) than
introductory and terminal elements, but experienced less blurring. It would be reasonable
to predict that trill elements would experience less blurring, given that trill elements have
lower frequencies and are more tonal than introductory or terminal elements (Figure 1) and
therefore should experience less degradation (Brown and Hanford 2000; Slabbekoorn et al.
2002). Our results for the interaction between element and distance supported this predic-
tion; we observed little variation in the blurring of trill elements as transmission distance
increased for both sexes. In contrast, terminal elements showed a higher blur ratio than
did trill elements, and blur ratio increased with distance for terminal elements. However,
both males and females appeared to compensate for this by singing terminal elements that
had a higher signal-to-noise ratio (Table S2). By comparison, introductory elements fell in
between terminal and trill elements with regards to signal-to-noise ratio and blur ratio, but
male introductory elements experienced greater excess attenuation, while female introduc-
tory elements showed a greater tail-to-signal ratio; this suggests that trills are likely more
important for long-distance communication (given that they experience less degradation
over further distances, Barker et al. 2009), whereas introductory elements and terminal
elements are likely most important over shorter distances and potentially used by receivers
to locate individuals at closer ranges (Morton 1986). Additionally, these elements may aid
receivers in determining the signallers identity (Bee et al. 2001; Sandoval et al. 2014), given
that these components of the song are highly variable (unpublished data).

Male vs. female transmission

Sex of the signaller may play a role in the attenuation and degradation of animal signals,
but to date the Acoustic Adaptation Hypothesis has primarily been tested only on male
acoustic signals (Morton 1975; Boncoraglio and Saino 2007). The differences we found
between the sexes in the degradation of song elements (i.e. source population significantly
affected the signal-to-noise ratio of males vs. blur ration and excess attenuation for females)
may reflect differences in communication strategies between sexes (Langmore 1998). Given
that females tend to be less conspicuous than males when singing (females produce fewer
 20 B. A. Graham et al.

songs, and sing primarily from lower perches in the understory; Topp and Mennill 2008;
Barker and Mennill 2009), and that female songs degrade faster as distance increases (Barker
et al. 2009), this suggests that male songs and singing behaviour are likely better adapted
for transmitting longer distances than females (Barker and Mennill 2009; Barker et al.
2009). Furthermore, duetting is an important aspect of the vocal behaviour in this spe-
cies (Mennill and Vehrencamp 2005), and while the average distance between pairs when
performing duets is approximately 20m, the majority of duets are produced between 0
and 10m (Mennill and Vehrencamp 2008). These observations suggest that female signals
are not adapted to maximize transmission distance but rather optimized to communicate
through dense vegetation over shorter distances with their breeding partners, especially
since female song is known to play a role in coordinating breeding activities (Ritchison
1983; Sonnenschein and Rayer 1983). At all three sites, we have observed females produc-
ing songs and calls at the nest during nest building, and from the nest while incubating
eggs or brooding young (Kovach 2013). Given that females vocalize so much near or at
the nest, they risk drawing the attention of potential predators from afar. Therefore, female
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signals may be quieter and experience greater degradation with increasing distance because
broadcasting loud far-reaching signals could be detrimental to their fitness. The Acoustic
Adaptation Hypothesis often assumes that animal vocalizations are adapted to maximize
transmission range while minimizing degradation (Boncoraglio and Saino 2007); however,
differences in the transmission properties of males and females (Barker et al. 2009) may
reflect different life history traits.
Past studies have emphasized the role that culture has on the evolution of songs through
forces that include selection, learning biases and drift (Lynch 1996; Podos and Warren
2007). Importantly, song transmission properties may affect learning, especially in the light
of a recent study by Peters et al. (2012) that suggested young birds preferentially learn the
least-degraded songs. As mentioned previously, terminal elements at both our wet forest
and montane forest sites tend to be more tonal (e.g. the third through fifth male terminal
elements in the second row of Figure 1) than at our dry forest site, where birds use terminal
elements with sharp rising or falling frequency sweeps (e.g. the second male terminal ele-
ment in the second row of Figure 1). Differences in the transmission properties of different
element types could explain element differences among our three sites; ongoing research
will explore differences in elements among these and other sites.

Conclusion
Our study does not suggest that acoustic variation among the three populations of Rufous-
and-white Wrens has been driven heavily by acoustic adaptation to three different tropical
forest environments. While previous research makes it clear that these birds songs are
adapted for transmission through forests vs. fields (Barker 2008), our current work does not
suggest that they are specifically adapted to different types of forest. In our study, playback
site (in particular ambient noise) played an important role in the transmission and degra-
dation of both male and female elements. In contrast, source population had a weak effect
on the degradation of elements for both males and females. Furthermore, the interaction
between playback site and source population did not suggest local adaptation, given that
song elements did not transmit better at their respective sites. While male and female ele-
ments showed similar patterns of degradation, we did observe a few important differences.
 Bioacoustics 21

For example, male elements appeared to be optimized to transmit most efficiently through
their environment, given that we found elements recorded from populations where ambi-
ent noise is higher had a higher signal-to-noise ratio. In contrast, female elements showed
no differences in signal-to-noise ratio among sites. However, we did observe that source
population affected blur ratio and excess attenuation of female elements. Elements recorded
from the montane forest site (the habitat with the highest vegetation density) had a lower
blur ratio, suggesting that these elements are optimized for transmission through densely
vegetated habitat. While our observations of male and female elements do not suggest local
adaptations, they may be indicative of plastic modifications, but further studies are neces-
sary to support this idea. Importantly, this study emphasizes the transmission differences
between sexes, which is likely reflective of behavioural and life history differences between
sexes. Whereas male song elements are likely maximized for long-range transmission, this
does not seem to be the case for female songs; female song elements seem to be optimized
for transmission through dense vegetation. This is important given that females often sing
from the densely vegetated understory and will also sing songs when they are concealed in
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their nests. Future studies should continue to compare male and female songs and singing
strategies to not only increase our understanding of the function of female song (Riebel
2003) but to better understand the behaviour and ecology of birds overall.

Acknowledgements
We thank N. Rehberg-Besler for field assistance. We thank R. Blanco and the Area de Conservacion
Guanacaste and F. Comacho and the University of Georgia Costa Rica Campus for assistance and
logistical support.

Disclosure statement
No potential conflict of interest was reported by the authors.

Funding
Funding was provided by an Ontario Graduate Scholarship (OGS), a Queen Elizabeth II Graduate
Scholarship in Science and Technology, a Chapman Grant from the American Museum of Natural
History, a Student Research Grant from the Animal Behaviour Society and an Alexander Wetmore
Research Award from the American Ornithologist Union to B.A.G.; funding was provided by the
Ministerio de Ciencia y Tecnologa (MICIT),the Consejo Nacional para Investigaciones Cientficas
y Tecnolgicas (CONICIT) and OGS to L.S.; and grants from the Natural Sciences and Engineering
Research Council of Canada (NSERC), the Canada Foundation for Innovation (CFI), the Government
of Ontario and the University of Windsor to D.J.M.

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